GASEOUS

STERILISATION
 STERILISATION
Elimination/ killing all forms of living or
biological agents
Types of sterilisation
 Dry & moist heat
 Filtration
 Radiation
 Gaseous
Demerits of heat sterilisation
 Thermolabile solid medicaments: precipitation
or crystallisation
 Thermolabile equipment: articles of plastic,
electrical diagnostic equipment, delicate rubber
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items
 GASEOUS STERILISATION
Sterilisation with chemical in the gaseous state
is a method of greater application.
Ethylene oxide
General properties
 Simplest cyclic ether
 Colourless gas
 Ethereal odour
 Liquefied easily
 Boils at approx. 10.8°C
 Dissolves in water and organic solvents
 Concentrations greater than 3% in air are highly
inflammable
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 ETHYLENE OXIDE
High inflammability can be overcome by:
 Admixture with an Inert Gas:
 Ethylene oxide and carbon dioxide or
halogenated hydrocarbons (1:9) are safe with
air.
 Cryoxcide - mixture of ethylene oxide 11%,
trichlorofluoromethane 79% and
dichlorodifluoromethane 10% .
 Sterethox – ethylene oxide 12% and
dichlorodifluoromethane 88%.

Carbon dioxide is less popular because of its high vapour

pressure. Less ethylene oxide is present in the vapour
when
carbon dioxide is used.
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 ETHYLENE OXIDE
 Using the Gas in the Absence of Oxygen: Sterilisation
carried out at sub atmospheric pressure. Pure or mixture
(90% ethylene oxide & 10% CO2) of ethylene oxide is used
in chamber from which at least 95% air has been removed.
Carbon dioxide reduces but not eliminates the
inflammability,
therefore should be used in specially designed equipment.

Mechanism of Action
 Alkylating agent. Alkyl groups are firmly bound, hence,
bactericidal effect.
 Active against all microorganisms, vegetative bacteria
and spores.
 Most resistant: Bacillus subtils var. globigii and var.
niger spores are used for testing ethylene oxide
sterilisers.
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 FACTORS INFLUENCING
EFFICIENCY

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FACTORS INFLUENCING EFFICIENCY
Concentration
 Expressed in mg/l – sterilisation rate depends on
partial pressure of ethylene oxide, which is
determined by amount in specified volume of
chamber atmosphere.
 Concentration for sterilisation range from 200 to
1000 mg/litre. Since dilution coefficient is 1, if
concentration is doubled - exposure time is halved.
Exposures required to kill 5 ×106 spores of
Bacillus subtilis
ETHYLENE OXIDEvar. globigii at 25°C
EXPOSURE TIME (h)
CONCENTRATION (mg/litre)

88 10
442 4
7 884 2
 FACTORS INFLUENCING
EFFICIENCY
Temperature
 Sterilisation achieved in lesser time with rise in
temperature.
 Within 5 to 40°C the sterilising dose (i.e. product of time
and concentration) can be halved for each increase of
17°C.
 For thermolabile materials; 60°C is fixed as the upper limit.

Relative humidity
 ‘Some moisture is necessary but a little is better than a lot.’
 Deficiency of water in the cells prevents ionisation of
groups susceptible to alkylation.
 At high humidities, the ethylene oxide in the cells is diluted
to below the most effective concentration.
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 FACTORS INFLUENCING
EFFICIENCY
Relative humidity
 Dehydrated organisms are difficult to destroy because:
Proteins contain linkages with H O molecules
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If dehydration takes place; these bonds replaced by
stronger links
Not available for combination with ethylene oxide
Organisms become resistant

 Sufficient moisture must be present –

 During exposure (40-50%). Water molecules help
carry ethylene oxide through plastic films of polar
nature (nylon & cellulose) while ethylene oxide assists
permeation of water through non polar polyethylene.
 During the pre-vacuum period: Resistant organisms
produced by vacuum dehydration can be restored to
normal sensitivity only by complete wetting or
9 exposure to near-saturation humidity for several days.
 FACTORS INFLUENCING
EFFICIENCY
Physical Protection of Organisms
• Articles for gaseous sterilisation must be clean, else,
organisms may enclose in gas impermeable deposits.
• Films of organic matter (blood or serum), less
protection than films containing crystals (dried
nutrient broth).
• Spores remain viable inside crystals for long time.
• Ethylene oxide alone – inadvisable for thermolabile
powders and crystals (spores inside the crystals
remain viable).
• Surface sterilisation of talc by ethylene oxide is
adequate because it is insoluble in body and internal
contamination of the particles of a mined insoluble
10 substance is highly improbable.
 FACTORS INFLUENCING
EFFICIENCY
Power of Penetration
 Good ability of penetration (paper, fabrics, plastic,
rubber)
 Post-sterilisation aseptic handling is eliminated.
 Suitable for:
 Powders in rubber-capped vials
 Talc in bulk
 Electrical diagnostic equipments
 Eye-drop bottles
 Blankets
 Residual gas escapes readily from inside wrappers
after sterilisation.
 Use only those plastics which are permeable to:
 Ethylene oxide: polyesters are imperpeable
 Air : to reduce risk of bursting during sterilisation
 Water vapour : ensure satisfactory humidity
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conditions
 FACTORS INFLUENCING
EFFICIENCY
Absorption
Many materials absorb ethylene oxide; effects:
 Reduction of the gas concentration in the chamber
atmosphere.
 Since gas is toxic, sterilised articles cannot be used until
absorbed gas has escaped. Method of desorption is
‘airing’.
 Article receives an additional exposure during desorption.

Toxicity
 The inhalation toxicity approximates to that of ammonia.
 May cause irritation, nausea and dizziness
 Of concern is the vesicant action of the moist compound.
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 APPLICATIONS
 POWDERS
Thermolabile: Restricted to sterilisation of the surfaces of
substances Highly
reactive; must be examined for deterioration.
Thermostable : Bulk powders (talc and Sterilisable Maize Starch B.P.)
 EYE DROPS IN PLASTIC UNI-DOSE CONTAINERS
 BLANKETS and beddings in hospital wards
 EXTERIOR OF AMPOULES ethylene oxide may penetrate; not
recommended.
 EQUIPMENT
 Catheters, syringes, needles, i.v. sets, bottles, vials, blood
oxygenators.
 Some plastics such as polystyrene become crazed; so mixture of
ethylene oxide and Arcton(s) is used.
 Fragile rubber articles such as, surgeons gloves survive more
treatments by ethylene oxide than steam.
 Electro-diagnostic: Cystoscopes, bronchoscopes, ophthalmoscopes,
Geiger-Muller counters etc.
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 ADVANTAGES
 Suitable for thermolabile substances.
 Does not damage moisture-sensitive substances.
 Can be used for pre-packed articles because of
great penetrating power of ethylene oxide.
 Only few materials are damaged during process.
 Used in closed screen to increase the reliability of
sterility testing.

DISADVANTAGES
 Slow; unsuitable in emergencies
 Large batches require very expensive, sterilisers;
need skilled and regular maintenance.
 Expensive
 Hazards of inflammability; toxic
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 Formaldehyde
 Alkylating agent
 Used for fumigation of empty rooms after infectious
diseases.
 Inferior to ethylene oxide because:
a. Its penetrating power is weak.
b. High concentrations are difficult to maintain in the
atmosphere.
c. A high humidity is necessary.
d. It is readily inactivated.
e. Adsorbed gas is very difficult to remove.
f. It is very irritating to the respiratory tract.
g. The pure substance cannot be kept at ordinary
temperatures.
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